FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2899573458> ?p ?o ?g. }

• W2899573458 endingPage "204" @default.
• W2899573458 startingPage "184" @default.
• W2899573458 abstract "Enzymes are attractive tools for synthetic applications. To be viable for industrial use, enzymes need sufficient stability towards the desired reaction conditions such as high substrate and cosolvent concentration, non-neutral pH and elevated temperatures. Thermal stability is an attractive feature not only because it allows for protein purification by thermal treatment and higher process temperatures but also due to the associated higher stability against other destabilising factors. Therefore, high-throughput screening (HTS) methods are desirable for the identification of thermostable biocatalysts by discovery from nature or by protein engineering but current methods have low throughput and require time-demanding purification of protein samples. We found that nanoscale differential scanning fluorimetry (nanoDSF) is a valuable tool to rapidly and reliably determine melting points of native proteins. To avoid intrinsic problems posed by crude protein extracts, hypotonic extraction of overexpressed protein from bacterial host cells resulted in higher sample quality and accurate manual determination of several hundred melting temperatures per day. We have probed the use of nanoDSF for HTS of a phylogenetically diverse aldolase library to identify novel thermostable enzymes from metagenomic sources and for the rapid measurements of variants from saturation mutagenesis. The feasibility of nanoDSF for the screening of synthetic reaction conditions was proved by studies of cosolvent tolerance, which showed protein melting temperature to decrease linearly with increasing cosolvent concentration for all combinations of six enzymes and eight water-miscible cosolvents investigated, and of substrate affinity, which showed stabilisation of hexokinase by sugars in the absence of ATP cofactor. ENZYMES: Alcohol dehydrogenase (NADP+ ) (EC 1.1.1.2), transketolase (EC 2.2.1.1), hexokinase (EC 2.7.1.1), 2-deoxyribose-5-phosphate aldolase (EC 4.1.2.4), fructose-6-phosphate aldolase (EC 4.1.2.n)." @default.
• W2899573458 created "2018-11-16" @default.
• W2899573458 creator A5045914500 @default.
• W2899573458 creator A5059041911 @default.
• W2899573458 creator A5066001844 @default.
• W2899573458 creator A5069786286 @default.
• W2899573458 creator A5070830522 @default.
• W2899573458 creator A5085182693 @default.
• W2899573458 date "2018-12-03" @default.
• W2899573458 modified "2023-10-12" @default.
• W2899573458 title "nanoDSF as screening tool for enzyme libraries and biotechnology development" @default.
• W2899573458 cites W1497863497 @default.
• W2899573458 cites W1807286889 @default.
• W2899573458 cites W1817888094 @default.
• W2899573458 cites W1845181906 @default.
• W2899573458 cites W1943327251 @default.
• W2899573458 cites W1947975057 @default.
• W2899573458 cites W1974288263 @default.
• W2899573458 cites W1977877548 @default.
• W2899573458 cites W1983686919 @default.
• W2899573458 cites W1984145455 @default.
• W2899573458 cites W1984180361 @default.
• W2899573458 cites W1986098920 @default.
• W2899573458 cites W1995053693 @default.
• W2899573458 cites W1997642309 @default.
• W2899573458 cites W2002456691 @default.
• W2899573458 cites W2003468739 @default.
• W2899573458 cites W2011739595 @default.
• W2899573458 cites W2021928977 @default.
• W2899573458 cites W2029028635 @default.
• W2899573458 cites W2032556503 @default.
• W2899573458 cites W2040203327 @default.
• W2899573458 cites W2041240739 @default.
• W2899573458 cites W2047132220 @default.
• W2899573458 cites W2049061140 @default.
• W2899573458 cites W2052170790 @default.
• W2899573458 cites W2052708596 @default.
• W2899573458 cites W2054328264 @default.
• W2899573458 cites W2060194574 @default.
• W2899573458 cites W2064127935 @default.
• W2899573458 cites W2079029484 @default.
• W2899573458 cites W2079214535 @default.
• W2899573458 cites W2084039289 @default.
• W2899573458 cites W2099000443 @default.
• W2899573458 cites W2106246537 @default.
• W2899573458 cites W2109441358 @default.
• W2899573458 cites W2111817477 @default.
• W2899573458 cites W2114160655 @default.
• W2899573458 cites W2116888800 @default.
• W2899573458 cites W2118197578 @default.
• W2899573458 cites W2121351614 @default.
• W2899573458 cites W2132417094 @default.
• W2899573458 cites W2142690324 @default.
• W2899573458 cites W2143550120 @default.
• W2899573458 cites W2145147476 @default.
• W2899573458 cites W2148286031 @default.
• W2899573458 cites W2148874377 @default.
• W2899573458 cites W2161852363 @default.
• W2899573458 cites W2168239566 @default.
• W2899573458 cites W2211873152 @default.
• W2899573458 cites W2326306194 @default.
• W2899573458 cites W2407522898 @default.
• W2899573458 cites W2413316678 @default.
• W2899573458 cites W2530058745 @default.
• W2899573458 cites W2551031656 @default.
• W2899573458 cites W2558698854 @default.
• W2899573458 cites W2604295418 @default.
• W2899573458 cites W2613845589 @default.
• W2899573458 cites W2762609780 @default.
• W2899573458 cites W2763497884 @default.
• W2899573458 cites W2807574667 @default.
• W2899573458 cites W4210980745 @default.
• W2899573458 cites W188593863 @default.
• W2899573458 doi "https://doi.org/10.1111/febs.14696" @default.
• W2899573458 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7379660" @default.
• W2899573458 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30414312" @default.
• W2899573458 hasPublicationYear "2018" @default.
• W2899573458 type Work @default.
• W2899573458 sameAs 2899573458 @default.
• W2899573458 citedByCount "76" @default.
• W2899573458 countsByYear W28995734582019 @default.
• W2899573458 countsByYear W28995734582020 @default.
• W2899573458 countsByYear W28995734582021 @default.
• W2899573458 countsByYear W28995734582022 @default.
• W2899573458 countsByYear W28995734582023 @default.
• W2899573458 crossrefType "journal-article" @default.
• W2899573458 hasAuthorship W2899573458A5045914500 @default.
• W2899573458 hasAuthorship W2899573458A5059041911 @default.
• W2899573458 hasAuthorship W2899573458A5066001844 @default.
• W2899573458 hasAuthorship W2899573458A5069786286 @default.
• W2899573458 hasAuthorship W2899573458A5070830522 @default.
• W2899573458 hasAuthorship W2899573458A5085182693 @default.
• W2899573458 hasBestOaLocation W28995734581 @default.
• W2899573458 hasConcept C104317684 @default.
• W2899573458 hasConcept C109129352 @default.
• W2899573458 hasConcept C143065580 @default.
• W2899573458 hasConcept C147816474 @default.
• W2899573458 hasConcept C181199279 @default.

• next